Integrated Evaluation of Protective Effects of Cucurbita ficifolia Seed Extract on Systemic Biochemicals in a Dihydrotestosterone Estradiol Valerate Induced Benign Prostatic Hyperplasia in Rats

1. Introduction

Benign prostatic hyperplasia (BPH) is one of the most prevalent urological disorders affecting aging men and represents a major cause of lower urinary tract symptoms (LUTS) worldwide. Histologically, BPH is characterized by hyperplasia of stromal and epithelial cells within the transitional zone of the prostate, leading to progressive gland enlargement and bladder outlet obstruction. Epidemiological data indicate that histological evidence of BPH is present in approximately 50% of men by the age of 60 years and increases to nearly 90% in men over 80 years, underscoring its strong association with aging [1]. 

The pathophysiology of BPH is multifactorial and involves complex interactions among hormonal imbalance, chronic inflammation, oxidative stress, and growth factor signaling [2]. Androgens, particularly dihydrotestosterone (DHT), play a central role in prostatic growth by binding to androgen receptors and promoting cellular proliferation while inhibiting apoptosis. Estrogens further modulate this process by altering androgen receptor expression and stromal epithelial interactions. Experimental induction of BPH using combined dihydrotestosterone and estradiol valerate administration closely mimics the hormonal milieu observed in aging men and is widely accepted as a reliable animal model for studying BPH pathogenesis and therapeutic interventions [3,4]. 

Current pharmacological treatments for BPH include α-adrenergic blockers and 5α-reductase inhibitors such as finasteride and dutasteride [5]. While effective, these agents are often associated with adverse effects including sexual dysfunction, dizziness, hypotension, and reduced treatment adherence, particularly with long-term use. Consequently, there has been growing interest in alternative and complementary therapies, especially plant-derived compounds, that may offer comparable efficacy with improved safety profiles [4]. 

Cucurbita ficifolia (fig-leaf gourd), a member of the Cucurbitaceae family, has been traditionally utilized for its nutritional and medicinal properties. The seeds are rich in phytosterols, polyunsaturated fatty acids, antioxidants, and trace elements, which have been reported to exert anti-inflammatory, antioxidant, and hormone-modulating effects. Extracts from Cucurbita species have demonstrated beneficial effects in prostate health, including reduction of LUTS and inhibition of prostatic growth, although species-specific evidence for C. ficifolia remains limited [6,7]. 

Beyond localized prostatic changes, BPH and its experimental induction may influence systemic biochemical parameters, including serum electrolyte balance. Electrolytes such as sodium, potassium, calcium, chloride, and bicarbonate are essential for maintaining cellular homeostasis, neuromuscular excitability, and acid–base equilibrium. Hormonal dysregulation, oxidative stress, and renal functional changes associated with BPH may contribute to electrolyte disturbances; however, this aspect remains under-investigated [8]. 

Therefore, this study aimed to investigate the effect of methanol extract of Cucurbita ficifolia seeds on serum electrolyte profiles in a dihydrotestosterone estradiol valerate-induced benign prostatic hyperplasia model in male Wistar albino rats. To ahcieve the above aim, the following specific objectives were pursed. 

1. Divide the Rats into 5 groups of 6 rats each 
2. Determine the body and Organ weight of the experimental animals 
3. Induce Benign Prostatic hyperplasia in experimental animals 
4. Assessment of the prostate specific antigen (PSA) in both normal and BPH Animals 
5. Determine the concentration of Liver Enzymes- Alanine aminotransferase (ALT), Aspartate aminotransferase (AST) and Alkaline Phosphatase (ALP) 
6. Assess the Hematological Parameters- White blood cell (WBC), Red blood cell (RBC), Haemoglobin(Hb), Packed cell volume (PCV), Platelet count (PLT) 
7. Determine the lipid profiles of the rat 
8. Evaluate the Antioxidant Activities – Catalase (CAT), Superoxide dismutase (SOD), Reduced glutathione (GSH) and Malonaldehyde (MDA) 

 The findings are expected to provide experimental evidence supporting the systemic safety and therapeutic relevance of C. ficifolia seed extract in the management of BPH. 

1.1 Background Research

Despite extensive research on benign prostatic hyperplasia (BPH), current understanding remains largely centered on prostatic enlargement and lower urinary tract symptoms, with comparatively less attention given to the systemic biochemical consequences of the disease. Emerging evidence suggests that BPH is not merely a localized pathological condition but may involve broader metabolic and physiological disturbances that extend beyond the prostate [5,3]. Alterations in renal function and electrolyte balance are increasingly being recognized in association with bladder outlet obstruction and hormonal dysregulation. Disruptions in sodium, potassium, chloride, and bicarbonate homeostasis may arise due to impaired renal perfusion and altered tubular reabsorption [8,9]. Oxidative stress and inflammation are well-established contributors to BPH progression and are closely linked with systemic biochemical alterations such as lipid metabolism and endocrine dysfunction [8,12]. However, their interplay with electrolyte regulation remains insufficiently characterized.

Current pharmacological interventions, including 5α-reductase inhibitors and α-adrenergic blockers, primarily target androgen metabolism and smooth muscle tone but do not adequately address metabolic and oxidative disturbances [13]. Plant-derived bioactive compounds have gained increasing attention due to their ability to modulate multiple biological pathways simultaneously [4].

Although Cucurbita species have demonstrated beneficial effects on prostate health, there is a paucity of data regarding the specific effects of C. ficifolia, particularly in relation to systemic biochemical alterations such as electrolyte imbalance and renal function [6,7]. This study was therefore designed to investigate the effect of methanol extract of Cucurbita ficifolia seeds on electrolyte balance and associated biochemical parameters in a hormone-induced model of benign prostatic hyperplasia, with the aim of providing novel insights into its potential multi-system therapeutic benefits.

2. Research Methodology

2.1 Materials and Instrumentation

Dihydrotestosterone (DHT), Estradiol valerate (ESV), Potassium Chloride (Kcl), Methanol, and others not listed were purchased from Sigma-Aldrich, St. MO, USA. Commercial Olive oil (Goya foods, Jersey City, NJ, USA ) purchased from a local supermarket served as the vehicle for the preparation and administration of Dihydrotestosterone (DHT) and Estradiol valerate (ESV), All chemicals used for this research were of analytical grade. Biochemical assay kits were purchased from Randox Laboratories.

2.2 Plant Collection and Extraction

Fresh seeds of Cucurbita ficifolia (also known as Seven-Year Melon or fig-leaf gourd) were collected from New Market, Aba, Abia State, Nigeria, and authenticated at the Department of Forestry, College of Natural Resources and Environmental Management, Michael Okpara University of Agriculture, Umudike. The seeds were washed, air-dried under shade for two weeks, and pulverized into fine powder. The powdered material (600 g) was extracted with 80% methanol using the cold maceration method for 48 hours with occasional stirring. The extract was filtered and concentrated using a rotary evaporator at 40 °C and stored in an airtight container at 4 °C until use

2.3 Experimental Design:

Adult male rats weighing 180 -220g were obtained from the animal house, Faulty of veterinary Medicine, University of Nigeria, Enugu State, Nigeria. The rats were acclimatized for two weeks under standard laboratory conditions before the commencement of the experiment. The albino rats were randomly assigned into five groups of Six animals each: Olive oil was used as a vehicle for hormonal agents. The volume and dosage used was based on earlier reported experimental BPH protocols and established pharmacological activities of the respective agents. The use of olive oil as the hormonal vehicle and the administrative volume per rat every other dayfor 28 days have been previously reported by [14].

• Group I (Normal control): received olive oil (5 ml/kg body weight) orally daily and subcutaneous injection of olive oil (0.1ml per rat ) every other day.
• Group II (Disease control): received subcutaneous injections of 9 mg/kg body weight of dihydrotestosterone and 0.9 mg/kg body weight of estradiol Valerate every other day, and olive oil (0.1ml per rat ) orally daily for 28 days.
• Group III (Drug control): received DHT and estradiol Valerate as in Group II, and oral administration of finasteride (5 mg/kg) daily for 28 days.
• Group IV (Treatment I): received DHT and estradiol Valerate as above and 200 mg/kg body weight of C. ficifolia seed extract orally daily for 28 days.
• Group V (Treatment II): received DHT and estradiol Valerate and 400 mg/kg body weight of C. ficifolia seed extract orally daily for 28 days.

At the end of the 28 days, the rats were starved overnight and were euthanized by cervical dislocation and blood was collected by cardiac puncture into plain sample tube and allowed to clot.

Serum was separated within one hour of blood clotting by centrifugation at 3000xg for about 20mins after which they were collected using Pasteur pipette. The serum samples were stored in a refrigerator until required for use. The serum samples were used for the assay of serum electrolyte (sodium, potassium, bicarbonate, calcium and chloride). The organs (prostate, testis, liver, kidney and heart) were excised, rinsed in ice- cold 1.15% KCL, blotted and weighed using an electronic weighing balance (Ohanus Corporation, USA).

2.4 Induction of Benign Prostatic Hyperplasia

Benign prostatic hyperplasia (BPH) was induced in male Wistar rats using a combination of dihydrotestosterone (DHT) and estradiol Valerate (ESV) in a ratio of 10:1, as previously described with slight modifications. Rats received subcutaneous injections of DHT (9 mg/kg body weight) and estradiol Valerate (0.9 mg/kg body weight) every other day for 28 days. The hormone stock solution was prepared by dissolving 1 g of DHT and 0.1 g of estradiol Valerate in 100 mL of olive oil. This method reliably produces prostatic enlargement and hormonal imbalance consistent with experimental BPH models. [15]

2.5 Determination of Morphological Parameters

Body weights of all animals were recorded daily throughout the experimental period. Percentage change in body weight was calculated at the end of the study. Following sacrifice, organs were excised, cleared of adhering tissues, and weighed to determine relative organ weights. Relative prostate weight was expressed as g/1000 g body weight, while relative liver, kidney, heart, and testis weights were expressed as g/100 g body weight using standard formulas [16].

2.6 Assay of Prostate-Specific Antigen (PSA)

Serum prostate-specific antigen (PSA) levels were determined using an enzyme-linked immunosorbent assay (ELISA) kit based on the solid-phase sandwich principle. Briefly, PSA in the sample was immobilized using a goat anti-PSA antibody coated on microtiter wells. A horseradish peroxidase-conjugated monoclonal anti-PSA antibody was then added to form an antibody–antigen–enzyme complex. After washing to remove unbound components, tetramethylbenzidine (TMB) substrate was added, and color development was measured spectrophotometrically at 450 nm. PSA concentration was directly proportional to absorbance intensity [16, 17].

3.0 Determination of Antioxidant Levels:

Antioxidant enzyme activities (SOD, CAT, GPx) and lipid peroxidation (MDA) were measured using validated spectrophotometric assays in rat serum and tissue, as described in recent preclinical studies [18, 19].

3.1 Lipid Parameters

Serum total cholesterol, triglycerides, HDL‑C, and LDL‑C were quantified using standard enzymatic colorimetric methods adapted for rodent models, consistent with recent biochemical assessments in rodent nutrition studies [20].

3.2 Haematology

Complete blood count parameters (RBC, WBC, Hb, PCV, PLT) were determined using automated haematology analyzers, following protocols recently applied in rat models evaluating systemic biochemical effects [21].

3.3 Renal Function Tests

Serum creatinine, urea, and electrolyte concentrations were assessed using established enzymatic and ion‑selective electrode techniques in rats, as recently reported in 2025–2026 experimental work [21].

3.4 Liver Function Tests:

Serum ALT, AST, ALP, albumin, and total protein were measured using c olorimetric and enzymatic assays, consistent with the analytic methods described in recent rodent liver function studies [21].

3.5 Statistical analysis

The Statistical Package for the Social Sciences (SPSS, version 25.0, IBM Corp., Armonk, NY, USA) was used for data analysis. Data were expressed as mean ± standard Deviation (SD), n =6 per group. Statistical significance was determined using one-way analysis of variance (ANOVA) followed by Turkey’s Honestly Significant Difference(HSD), post hoc test. Mean with different superscript letters within the same column differ significantly. Comparison was made between the control and test groups. A value of p < 0.05 was considered statistically significant.

4. Results and Discussion

Benign prostatic hyperplasia (BPH) is a progressive, age-associated urological disorder characterized by non-malignant enlargement of the prostate gland and driven by complex interactions among androgen estrogen imbalance, oxidative stress, chronic inflammation, metabolic dysregulation, and altered growth factor signaling [22]. In the present study, combined administration of dihydrotestosterone (DHT) and estradiol Valerate successfully induced BPH in male wistar rats, as evidenced by increased prostate weight, elevated prostate-specifi c antigen (PSA), hormonal disturbances, oxidative stress, dyslipidaemia, renal dysfunction, and electrolyte imbalance [29]. Treatment with methanol extract of Cucurbita ficifolia seeds significantly ameliorated these pathological alterations, indicating a broad protective role against BPH-associated systemic complications.

Table 4.1: Effect of Curcubita ficifolia Methanol Extract on Relative Organ Weight of Dihydrotestosterone, Estradiol Valerate Induced BPH in Rats

Results are expressed as mean ± standard deviation, N=6. followed by Tukey HSD post hoc test. Mean values having different alphabets (a-c) as superscript down the column are significantly (p<0.05) different from each other.

The results of Table 4.1 show the effect of Cucurbita ficifolia on relative organ weight. The result showed that no significant change was observed in the weight of liver and testes (p>0.05) in all the groups. The kidney weights were significantly higher (p<0.05) in finasteride control group and group treated with 400mg/kg bwt of the extract compared to other group. Induction of benign prostatic hyperplasia with dihydrotestosterone (DHT) and estradiol valerate resulted in a significant increase (p<0.05) in relative weight of the prostate in BPH control group compared with the normal control group. Treatment with the extract of Cucurbita ficifolia produced a dose dependent reduction in weight of the prostate compared with the disease control group. The reduction was statistically significant (p<0.05) at the medium and high doses of the extract. Rats treated with finasteride also showed a significant decrease (p<0.05) in prostate weight relative to untreated BPH control group. The reduction observed in the high dose extract group was comparable to that of finasteride treated group. The significant increase in relative prostate weight observed in the disease control group confirms successful induction of prostatic hyperplasia. Prostate enlargement in BPH results primarily from androgen-driven stromal and epithelial proliferation, particularly mediated by dihydrotestosterone acting through androgen receptors [13,23]. Administration of C. ficifolia extract produced a dose-dependent reduction in prostate weight, with the higher dose yielding values comparable to the normal control. This finding suggests that the extract may attenuate androgen-mediated prostatic growth. Importantly, the absence of significant changes in liver and testes weights indicates that the extract did not exert overt systemic or reproductive toxicity

Table 4.2: Effect of Cucurbita ficifolia Methanol Extract on Prostate Specific Antigen (PSA) of Dihydrotestosterone -Estradiol Valerate Induced BPH in Rats

Results are expressed as mean ± standard deviation, N=6, followed by Tukey HSD post hoc test. Mean values having different alphabets (a-c) as superscript down the column are significantly (p<0.05) different from each other.

The results of Table 4.2 show the effect of Cucurbita ficifolia on prostate-specific antigen (PSA). PSA levels were significantly increased (p < 0.05) in the disease control group compared to the normal and drug control groups. Treatment with C. ficifolia extract significantly reduced PSA levels compared with the disease control. Elevated PSA levels are closely associated with prostatic epithelial activity, inflammation, and androgen receptor signaling. The marked increase in PSA observed in the disease control group is consistent with previous reports linking BPH progression to increased PSA expression [14]. Treatment with C. ficifolia extract significantly reduced PSA levels, particularly at the lower dose, indicating suppression of prostatic hyperactivity. This effect may be attributed to phytosterols and phenolic compounds present in Cucurbita species, which have been reported to inhibit 5α-reductase activity and modulate androgen signaling [6,4].

The results of Table 4.3 show the effect of Curcubita ficifolia on hematological parameters. The result showed no significant differences (p>0.05) in RBC, PCV, and Hb levels in all the groups. However, significant changes were seen in WBC and Platelet (PLT) counts. The disease control group showed a significant increase in WBC count compared to normal group. Platelet counts significantly decreased in disease control group compared to normal group. This indicates that neither BPH induction nor treatment with Curcubita ficifolia extract adversely affected erythropoiesis or caused anemia. However, significant changes were observed in WBC and platelet counts. The significant increase in WBC count in disease control group suggests an inflammatory or immune response associated with BPH induction [21].

Conversely, the significant decrease in platelet count may indicate altered platelet homeostasis during inflammation. Overall, the findings suggest that the plant extract possesses relative hematological safety despite mild inflammatory-related alterations.

Table 4.3: Effect of Curcubita ficifolia Methanol Extract on Hematological parameters of Dihdrotestosterone, Estradiol Valerate Induced BPH in Rats.

Results are expressed as mean ± standard deviation, N=6, followed by Tukey HSD post hoc test. Means values having different alphabets (a-c) as superscript down the column are significantly (p<0.05) different from each other.

Table 4.4: Effect of Curcubita ficifolia Methanol Extract on Liver function parameters of Dihydrotestosterone- Estradiol valerate Induced BPH in Rats

Results are expressed as mean ± standard deviation, N=6, followed by Tukey HSD post hoc test. Mean values having different alphabets (a-c) as superscript down the column are significantly (p<0.05) different from each other.

Based on the values in table 4.5, the liver enzyme parameters (ALT, AST and ALP) show that induction of BPH was associated with some significant elevation of liver injury markers (P< 0.05) compared to normal control group, Administration of Cucurbita ficifolia methanol extract produced partial improvement, particularly at 200 mg/kg for ALT/AST and at 400 mg/kg for ALP. Elevation of serum ALT and AST is generally associated with hepatocellular injury, whereas increased ALP is more commonly associated with hepatobiliary disturbances [31]. The reduction in the activities of these enzymes following treatment, particularly the reduction in ALP and the partial reduction in ALT and AST, may suggest a protective effect of C. ficifolia extract against the biochemical alterations associated with the induced disease condition. Plant-derived bioactive compounds, including flavonoids and other antioxidant constituents, have been reported to exert hepatoprotective effects through reduction of oxidative stress and attenuation of hepatocellular injury [32].

The results presented in Table 4.5 show variations in the serum total protein, albumin and globulin concentrations among the experimental groups. Serum total protein reflects the combined concentration of albumin and globulins and is influenced by hepatic protein synthesis, nutritional status, hydration and immune activity [33]. The relatively similar total protein concentrations between the normal control and disease control suggest that induction of the experimental BPH condition did not markedly disrupt overall serum protein concentration. The slight increase observed in the drug control may reflect treatment-related or physiological changes rather than a specific hepatic effect. The relatively lower albumin concentration in the disease control compared with the normal control may suggest a mild alteration in protein synthetic or metabolic processes. However, albumin has a relatively long half-life and is affected by several non-hepatic factors; therefore, a modest change should not by itself be interpreted as evidence of significant hepatic dysfunction. Overall, the relatively stable total protein concentrations, together with the moderate changes in albumin and globulin, suggest that the DHT/estradiol induced experimental condition did not cause a profound disturbance of serum protein status. The changes observed following C. ficifolia administration may reflect alterations in protein metabolism and/or inflammatory status rather than severe impairment of hepatic synthetic function=.

Table 4.5: Effect of Curcubita ficifolia Methanol Extract on renal function parameters of Dihydrotestosterone, Estradiol valerate Induced BPH in Rats.

Results are expressed as mean ± standard deviation, N=6, followed by Tukey HSD post hoc test. Mean values having different alphabets (a-c) as superscript down the column are significantly (p<0.05) different from each other.

The results of Table 4.5 showed the effect of Curcubita ficifolia extract on renal function parameters in Dihydrotestosterone, Estradiol Induced BPH in Rats. The diseased control group showed the highest serum urea and creatinin concentration compared with the normal control group. Treatment with Curcubita ficifolia extract at 200 and 400mg/kg reduced the serum urea and creatinine levels towards normal values. For serum electrolyte analysis, the disease control group exhibited lower sodium and chloride. Administion of with C. ficifolia extract restorede the sodium and chloride levels towards the normal values. Serum potassium and bicarbonate concentration shoed little variation among the experimental groups.

Renal function and electrolyte homeostasis were notably disrupted in BPH-induced rats. As shown in Table 4.5, the disease control group exhibited elevated creatinine levels, reduced sodium and chloride concentrations, and increased potassium levels. These changes may result from bladder outlet obstruction associated with BPH, leading to impaired renal perfusion and altered tubular handling of electrolytes [11, 8]. Treatment with C. ficifolia extract restored electrolyte levels toward normal and reduced creatinine concentration, indicating preservation of renal function. These findings suggest a nephroprotective effect of the extract, potentially mediated through antioxidant and anti-inflammatory mechanisms.

Table 4.6: Effect of Curcubita ficifolia Methanol Extract on Lipid profile of Dihydrotestostrone, Estradiol Valerate Induced BPH in Rats.

Results are expressed as mean ± standard deviation (SD), N=6, followed by Tukey HSD post hoc test Mean values having different alphabets (a-c) as superscript down the column are significantly (p<0.05) different from each other. The results of Table 4.6 show the effect of Curcubita ficifolia on serum lipid profile in Dihydrotestostrone, Estradiol Valerate Induced BPH in Rats. The disease control group showed a significant (p < 0.05) increase in total cholesterol , triacylglycerol, LDL-cholesterol , and VLDL-cholesterol compared with the normal control group, indicating disruption of lipid metabolism following BPH induction. HDL-cholesterol levels did not differ significantly among the experimental groups (p > 0.05).

Treatment with Cucurbita ficifolia methanol extract at 200 mg/kg and 400 mg/kg reduced total cholesterol, triacylglycerol, LDL-cholesterol, and VLDL-cholesterol compared with the disease control group. The reduction was more evident in the 200 mg/kg group for LDL-cholesterol, while both doses improved the overall lipid profile. Finasteride also significantly ameliorated the BPH-induced alterations in serum lipid parameters. The present study demonstrates that DHT- and estradiol valerate-induced BPH is associated with significant disturbances in lipid metabolism, as evidenced by elevated total cholesterol, triacylglycerol, LDL-cholesterol, and VLDL-cholesterol in the disease control group. These findings agree with previous reports that androgen-induced BPH is accompanied by dyslipidaemia and increased oxidative stress, both of which contribute to the progression of prostatic enlargement and inflammation [12,24].

Administration of Cucurbita ficifolia methanol extract improved the altered lipid profile by reducing serum total cholesterol, triacylglycerol, LDL-cholesterol, and VLDL-cholesterol. The hypolipidaemic effect of the extract may be attributed to its rich phytochemical constituents, including flavonoids, phenolic compounds, saponins, and other bioactive metabolites, which have been reported to inhibit lipid synthesis, enhance lipid metabolism, and reduce oxidative stress. These hypolipidaemic effects are likely attributable to phytosterols and polyunsaturated fatty acids abundant in Cucurbita seeds, which inhibit cholesterol absorption and modulate lipid metabolism [6].

Lower LDL and VLDL concentrations are particularly beneficial because these lipoproteins promote lipid accumulation, inflammation, and oxidative damage, all of which are implicated in BPH progression.

Interestingly, HDL-cholesterol remained relatively unchanged across all groups, suggesting that the extract exerted its lipid-lowering effect mainly by reducing atherogenic lipids rather than increasing protective HDL-cholesterol. Similar observations have been reported in experimental studies where plant-derived extracts improved serum cholesterol and triglycerides without significantly altering HDL concentrations.

The lipid-lowering effect observed in the extract-treated groups was comparable to that of the finasteride-treated group, suggesting that Cucurbita ficifolia possesses promising therapeutic potential against BPH-associated dyslipidaemia. The improvement in lipid homeostasis may contribute to the overall protective effect of the extract by reducing oxidative stress, inflammation, and prostatic tissue injury.

Table 4.7: Effect of Curcubita ficifolia Methanol Extract on Antioxidant parameters of Dihydrotesstosterone, Estradiol Vvalerate Induced BPH in Rats

Results are expressed as mean ± standard deviation, N=6, followed by Tukey HSD post hoc test. Mean values having different alphabets (a-c) as superscript down the column are significantly (p<0.05) different from each other. The results of Table 4.7 show the effect of Curcubita ficifolia on antioxidant parameters. The results showed that GPx, SOD, and CAT activities were significantly reduced, while MDA levels were significantly increased in the disease control group, indicating oxidative stress. Treatment with C. ficifolia extract restored antioxidant enzyme activities and significantly reduced MDA levels. Oxidative stress is a key pathogenic mechanism in BPH, contributing to cellular proliferation, stromal remodeling, and chronic inflammation. The significant reduction in antioxidant enzymes (SOD, CAT, and GPx) and increased malondialdehyde (MDA) levels observed in the disease control group confirm enhanced oxidative stress and lipid peroxidation. Treatment with C. ficifolia extract restored antioxidant enzyme activities and significantly reduced MDA levels, demonstrating potent antioxidant capacity. These findings indicate depletion of the endogenous antioxidant defense system and enhanced lipid peroxidation following hormonal induction of BPH.

Oxidative stress is increasingly recognized as an important contributor to the initiation and progression of BPH. Excessive production of reactive oxygen species (ROS) damages cellular lipids, proteins, and nucleic acids, promotes chronic inflammation, and stimulates abnormal proliferation of prostatic stromal and epithelial cells. Consequently, impairment of antioxidant enzymes such as GPx, SOD, and CAT contributes to disease progression [25].

Administration of Cucurbita ficifolia methanol extract significantly restored the activities of GPx, SOD, and CAT while reducing MDA concentration compared with the disease control group. These findings indicate that the extract effectively attenuated oxidative stress and protected the prostate against ROS-mediated damage. The observed antioxidant activity may be attributed to the presence of phytochemicals such as flavonoids, phenolic compounds, tannins, and terpenoids, which are known to scavenge free radicals, inhibit lipid peroxidation, and enhance endogenous antioxidant enzyme activities [24].

The significant reduction in MDA concentration following treatment further demonstrates that the extract inhibited lipid peroxidation and preserved membrane integrity. MDA is a well-established biomarker of oxidative damage, and elevated MDA levels have consistently been associated with BPH severity and chronic prostatic inflammation [28].. Therefore, the reduction in MDA observed in the extract-treated groups suggests effective protection against oxidative injury. These findings are consistent with reports that plant-derived antioxidants can suppress oxidative pathways involved in BPH progression [5].

Table 4.8: Effect of Curcubita ficifolia Methanol Extract on Reproductive Hormonal Level of Dihydrosterone, Estradiol Induced BPH in Rats

Results are expressed as mean ± standard deviation, N=6. Mean values having different alphabets (a-c) as superscript down the column are significantly (p<0.05) different from each other. The results of Table 4.8 show the effect of Curcubita ficifolia on reproductive hormone levels. The results showed that there were significant changes across all groups (p<0.05). The results show that there were significant increases in testosterone (16.23±0.40 ng/ml), FSH (7.45±0.42 miu/ml), and LH (5.47±0.29 miu/ml) in the disease control compared to the normal and drug controls. At 200 mg/kg, testosterone (14.40±0.19 ng/ml), FSH (6.30±0.14 miu/ml), and LH (4.84±0.19 miu/ml) were significantly lowered, while at the 400 mg/kg, they significantly decreased similar to the values of the controls.

Hormonal analysis revealed significant elevations in testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) in the disease control group, reflecting disruption of the hypothalamic–pituitary gonadal axis induced by exogenous hormone administration. Altered gonadotropic signaling has been associated with prostate enlargement and BPH progression [26]. Treatment with C. ficifolia extract significantly restored these hormonal levels toward normal, particularly at the higher dose, suggesting modulation of endocrine signaling particularly at the higher dose, suggesting modulation of endocrine signaling [29]. This hormonal regulation likely contributed to the reductions in prostate weight and PSA levels observed in treated groups [27].

Overall, the findings of this study demonstrate that methanol extract of Cucurbita ficifolia seeds exerts multi-target protective effects against BPH-induced pathological alterations. The extract improved prostate morphology, hormonal balance, antioxidant status, lipid metabolism, renal function, and electrolyte homeostasis. Unlike finasteride, which primarily targets androgen metabolism, C. ficifolia appears to act through complementary mechanisms, offering broader systemic benefits and supporting its potential as a safe adjunct or alternative therapeutic agent in the management of benign prostatic hyperplasia.

5. Conclusion

The present study demonstrates that methanol extract of Cucurbita ficifolia seeds effectively attenuates hormone-induced benign prostatic hyperplasia in Wistar rats. The extract significantly reduced prostate enlargement and prostate-specific antigen levels while restoring antioxidant defenses, improving lipid metabolism, preserving renal function, and normalizing electrolyte and hormonal homeostasis. These findings indicate that C. ficifolia exerts its protective effects through integrated mechanisms involving anti-androgenic, antioxidant, anti-inflammatory, and metabolic regulatory pathways. Importantly, the absence of adverse effects on vital organs suggests a favorable safety profile. Collectively, this study positions Cucurbita ficifolia as a promising multi-target phytotherapeutic agent for BPH management. However, further studies, particularly well-designed clinical trials, are required to validate its efficacy, elucidate its molecular mechanisms, and establish its translational potential in human populations.

Authors Contribution: Conceptualization and Methodology, ANK, OC and OCO., Validation, Formal analysis and Investigation, ANK, APO and OCO., Data Curation, APO and OOKL, and Writing Original draft preparation, ANK, OCO and SAC. Writing review and editing, ENU, MF, OOKL and CEC., Project Administration and Funding acquisition. ANK, ENU, OC, APO and MJE. Software, SAC, OBK, and OSO, Writing Original Draft Preparation ANK, OC and OCO, Visualization ANK, OO and MJE. All authors have read and agreed to published version of the manuscript.

Funding

This reseach receives no external funding.

Acknowledgements

The authors acknowledge the Department of Biochemistry at Michael Okpara University of Agriculture, Umudike, for providing the laboratory facilities for this research. Special thanks to the laboratory staff for their technical support during the animal experiments.

Conflict of Interest

The authors declare no conflict of interest related to this research.

Ethical Approval: All experimental procedures involving animals were conducted in compliance with institutional guidelines and were approved by the Animal Ethics Committee of Michael Okpara University of Agriculture, Umudike.

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